Performance of Loop-Mediated Isothermal Amplification for the Identification of Submicroscopic Plasmodium falciparum Infection in Uganda

Am J Trop Med Hyg. 2017 Dec;97(6):1777-1781. doi: 10.4269/ajtmh.17-0225. Epub 2017 Oct 5.


Accurately identifying and targeting the human reservoir of malaria parasitemia is critical for malaria control, and requires a reliable and sensitive diagnostic method. Loop-mediated isothermal amplification (LAMP) is increasingly used to diagnose submicroscopic parasitemia. Although most published studies report the sensitivity of LAMP compared with nested polymerase chain reaction (PCR) as ≥ 80%, they have failed to use a consistent, sensitive diagnostic as a comparator. We used cross-sectional samples from children and adults in Tororo, Uganda, a region with high but declining transmission due to indoor residual spraying, to characterize the sensitivity and specificity of pan-Plasmodium LAMP for detecting submicroscopic infections. We compared LAMP results targeting a mitochondrial DNA sequence conserved in all Plasmodium species, performed on DNA extracted from dried blood spots, to those of a gold standard quantitative PCR assay targeting the var gene acidic terminal sequence of Plasmodium falciparum (varATS qPCR), performed on DNA extracted from 200 µL of whole blood. Using LAMP and varATS qPCR increased the detection of parasitemia 2- to 5-fold, compared with microscopy. Among microscopy-negative samples, the sensitivity of LAMP was 81.5% for detecting infection ≥ 1 parasites/µL. However, low density infections were common, and LAMP failed to identify more than half of all infections diagnosed by varATS qPCR, performing with an overall sensitivity of 44.7% for detecting submicroscopic infections ≥ 0.01 parasites/µL. Thus, although the LAMP assay is more sensitive than microscopy, it missed a significant portion of the submicroscopic reservoir. These findings have important implications for malaria control, particularly in settings where low-density infections predominate.

MeSH terms

  • Adult
  • Child
  • Child, Preschool
  • Cross-Sectional Studies
  • DNA, Protozoan / isolation & purification
  • Female
  • Humans
  • Infant
  • Malaria, Falciparum / diagnosis*
  • Malaria, Falciparum / epidemiology*
  • Male
  • Middle Aged
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques / methods*
  • Plasmodium falciparum / isolation & purification*
  • Prevalence
  • Sensitivity and Specificity
  • Uganda / epidemiology


  • DNA, Protozoan